Three-dimensional structure of p21 in the active conformation and analysis of an oncogenic mutant.

The three-dimensional structure of the active guanosine triphosphate (GTP)-analogue-containing complex of the H-ras-encoded p21 has been determined. It was necessary to correct the topology of p21 as published earlier. The structure analysis shows all of the interactions between protein and GTP and how the important cofactor Mg2+ is bound. From the oncogenic mutants of p21 crystallized, a Gly12 to Arg mutation has been analyzed in detail. It shows that the overall structure of the mutant is not perturbed and that the side chain of Arg12 is coming close to the gamma-phosphate for an interaction.

[1]  Irving S. Sigal,et al.  Cloning of bovine GAP and its interaction with oncogenic ras p21 , 1988, Nature.

[2]  C. Marshall,et al.  The cytoplasmic protein GAP is implicated as the target for regulation by the ras gene product , 1988, Nature.

[3]  A. Gilman,et al.  G proteins: transducers of receptor-generated signals. , 1987, Annual review of biochemistry.

[4]  S H Kim,et al.  Three-dimensional structure of an oncogene protein: catalytic domain of human c-H-ras p21. , 1988, Science.

[5]  Frank McCormick,et al.  ras GTPase activating protein: Signal transmitter and signal terminator , 1989, Cell.

[6]  B. Clark,et al.  Structural details of the binding of guanosine diphosphate to elongation factor Tu from E. coli as studied by X‐ray crystallography. , 1985, The EMBO journal.

[7]  F. Eckstein,et al.  Nucleoside phosphorothioates. , 1970, Journal of the American Chemical Society.

[8]  W. Kabsch,et al.  Structure of the guanine-nucleotide-binding domain of the Ha-ras oncogene product p21 in the triphosphate conformation , 1989, Nature.

[9]  M. Barbacid.  ras genes. , 1987, Annual review of biochemistry.

[10]  B. Clark,et al.  Homologies in the Structures of G-Binding Proteins: An Analysis Based on Elongation Factor EF-TU , 1989, Bio/Technology.

[11]  K. Halliday.  Regional homology in GTP-binding proto-oncogene products and elongation factors. , 1983, Journal of cyclic nucleotide and protein phosphorylation research.

[12]  P. Seeburg,et al.  Biological properties of human c-Ha-ras1 genes mutated at codon 12 , 1984, Nature.

[13]  W. Merrick,et al.  GTP-binding domain: three consensus sequence elements with distinct spacing. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[14]  E. Pai,et al.  Crystallization and preliminary X-ray analysis of the human c-H-ras-oncogene product p21 complexed with GTP analogues. , 1989, Journal of molecular biology.

[15]  J. Knowles.  Enzyme-catalyzed phosphoryl transfer reactions. , 1980, Annual review of biochemistry.

[16]  A. Wittinghofer,et al.  Inhibition of GTPase activating protein stimulation of Ras-p21 GTPase by the Krev-1 gene product. , 1990, Science.

[17]  J. L. Bos,et al.  ras oncogenes in human cancer: a review. , 1989, Cancer research.

[18]  Hall Kr.  Regional homology in GTP-binding proto-oncogene products and elongation factors , 1983 .

[19]  F. Jurnak.  Structure of the GDP domain of EF-Tu and location of the amino acids homologous to ras oncogene proteins. , 1985, Science.